Assessment of hospital environmental and personal protective equipment contamination with influenza A and B viruses by ultrasensitive droplet digital PCR

Ningneng Zhai; Rongheng Tang; Ziyu Huang; Chenghan Wang; Yunhua Huang; Yan Long; Jiaqi Luo; Jinhua Qin; Yifang Huang

doi:10.1016/j.diagmicrobio.2025.116963

Assessment of hospital environmental and personal protective equipment contamination with influenza A and B viruses by ultrasensitive droplet digital PCR

Diagn Microbiol Infect Dis. 2025 Jun 23;113(3):116963. doi: 10.1016/j.diagmicrobio.2025.116963. Online ahead of print.

Authors

Ningneng Zhai¹, Rongheng Tang¹, Ziyu Huang², Chenghan Wang¹, Yunhua Huang¹, Yan Long¹, Jiaqi Luo¹, Jinhua Qin¹, Yifang Huang³

Affiliations

¹ Department of Clinical Laboratory, the First Affiliated Hospital of Guangxi Medical University, Key Laboratory of Clinical Laboratory Medicine of Guangxi Department of Education, Guangxi Medical University, Nanning 530021, China.
² Department of Pediatrics, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China.
³ Department of Clinical Laboratory, the First Affiliated Hospital of Guangxi Medical University, Key Laboratory of Clinical Laboratory Medicine of Guangxi Department of Education, Guangxi Medical University, Nanning 530021, China. Electronic address: 0501hyf@163.com.

PMID: 40609121
DOI: 10.1016/j.diagmicrobio.2025.116963

Abstract

Objectives: This study employed ultra-sensitive droplet digital PCR (ddPCR) to assess contamination of influenza A and B in hospital laboratories, patient wards, and personal protective equipment (PPE).

Methods: The analytical performance of ddPCR for detecting influenza A and B viruses was firstly assessed using virus transport medium (VTM) simulation samples and real simulation samples. Subsequently, a total of 22 high-touch surface samples were collected from the laboratory environment, along with five PPE samples from laboratory staff. Additionally, we collected 42 environmental surface samples from the wards of patients infected with influenza A and B, as well as ten PPE samples from health care workers (HCWs). All samples were analyzed using ddPCR.

Results: The results indicated that ddPCR can simultaneously detect the RNA of influenza A and B at a minimum concentration of 0.08 copies/µL in both VTM and real simulation samples, demonstrating exceptional stability and sensitivity. In the laboratory environment, influenza RNA was detected on 6 out of 22 high-touch surfaces, resulting in a positivity rate of 27.27 %. Furthermore, influenza RNA was detected on the laboratory staff's PPE. The overall positive rate of the wards housing influenza patients was 20 % (8/40); no influenza RNA was detected on two air outlet fans or on the PPE of the HCWs.

Conclusion: Ultra-sensitive ddPCR can effectively detect trace amounts of the influenza virus in the environment, including high-touch surfaces in laboratories, patient wards, and PPE. This advancement provides compelling evidence that this method is an innovative tool for epidemic prevention and control.

Keywords: Droplet digital PCR; Hospital environmental; Influenza A virus; Influenza B virus; Personal protective equipment.